2. Cut the membrane (Nitrocellulose or PVDF) and
6 pieces of blotting paper to the same size as the gel.

(optional) Mark the well
positions on the membrane with a pen or pencile.

3. For nitrocellulose membrane, wet the membrane in the transfer buffer.
For PVDF membrane, wet the membrane for 15 seconds in 100% methanol,
then soak for 2 minutes in water, then equilibrate the membrane for 5 minutes in the
transfer buffer.

5. Make sure no air bubbles get trapped between the gel and the membrane.

6. Insert the stack into a semi-dry or tank blotter as shown. For tank transfer,
sandwich the membrane/gel/filter paper between two sponges and insert into the cassette.
Fill the tank with the transfer buffer.